Annals of Microbiology

, Volume 68, Issue 4, pp 207–216 | Cite as

Identification and analysis of the function of surface layer proteins from three Lactobacillus strains

  • Jun Meng
  • Qiu-Xiang Zhang
  • Rong-Rong Lu
Original Article


To identify and investigate the role of surface layer proteins (SLPs) on the probiotic properties of Lactobacillus strains, SLPs were extracted from Lactobacillus bulgaricus fb04, L. rhamnosus fb06, L. gasseri fb07, and L. acidophilus NCFM by 5 mol/L lithium chloride. The molecular masses of the four SLPs were approximately 45–47 kDa as analyzed by SDS-PAGE. Hydrophobic amino acids were the main components of the four SLPs. The secondary structure content of the four SLPs showed extensive variability among different strains. After the SLPs were removed from the cell surface, the autoaggregation ability, coaggregation ability, and gastrointestinal tolerability of the four lactobacilli were significantly reduced as compared with the intact cells (P < 0.05). When exposed to bile salt stress, L. rhamnosus fb06, L. gasseri fb07, and L. acidophilus NCFM expressed more SLPs as determined by Bradford method. In conclusion, the four lactobacilli all possessed functional SLPs, which had positive contributions to the probiotic properties of the four Lactobacillus strains. This research could reveal the biological contributions of SLPs from Lactobacillus strains and offer a theoretical basis for the application of lactobacilli and their SLPs in food and pharmaceutical industries.


Surface layer proteins Lactobacillus Amino acid composition Secondary structure Probiotic function 



This study was supported by the grant from the National Natural Science Foundation of China (No. 31471696, 31701542) and Province Key Laboratory of Transformation and Utilization of Cereal Resource (PL2016009).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Åvall-Jääskeläinen S, Palva A (2005) Lactobacillus surface layers and their applications. FEMS Microbiol Rev 29:511–529CrossRefPubMedGoogle Scholar
  2. Beganović J, Frece J, Kos B, Leboš Pavunc A, Habjanič K, Šušković J (2011) Functionality of the S-layer protein from the probiotic strain Lactobacillus helveticus M92. Antonie Van Leeuwenhoek 100:43–53CrossRefPubMedGoogle Scholar
  3. Boot HJ, Kolen C, Van Noort JM, Pouwels PH (1993) S-layer protein of Lactobacillus acidophilus ATCC 4356: purification, expression in Escherichia coli, and nucleotide sequence of the corresponding gene. J Bacteriol 175:6089–6096CrossRefPubMedPubMedCentralGoogle Scholar
  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  5. Chen X, Xu J, Shuai J, Chen J, Zhang Z, Fang W (2007) The S-layer proteins of Lactobacillus crispatus strain ZJ001 is responsible for competitive exclusion against Escherichia coli O157: H7 and Salmonella typhimurium. Int J Food Microbiol 115:307–312CrossRefPubMedGoogle Scholar
  6. Collado MC, Meriluoto J, Salminen S (2008) Adhesion and aggregation properties of probiotic and pathogen strains. Eur Food Res Technol 226:1065–1073CrossRefGoogle Scholar
  7. Frece J, Kos B, Svetec I-K, Zgaga Z, Mrša V, Šušković J (2005) Importance of S-layer proteins in probiotic activity of Lactobacillus acidophilus M92. J Appl Microbiol 98:285–292CrossRefPubMedGoogle Scholar
  8. Gerbino E, Mobili P, Tymczyszyn E, Fausto R, Gómez-Zavaglia A (2011) FTIR spectroscopy structural analysis of the interaction between Lactobacillus kefir S-layers and metal ions. J Mol Struct 987:186–192CrossRefGoogle Scholar
  9. Golowczyc M, Mobili P, Garrote G, Abraham A, De Antoni G (2007) Protective action of Lactobacillus kefir carrying S-layer protein against Salmonella enterica serovar Enteritidis. Int J Food Microbiol 118:264–273CrossRefPubMedGoogle Scholar
  10. Golowczyc MA, Mobili P, Garrote GL, de los Angeles Serradell M, Abraham AG, De Antoni GL (2009) Interaction between Lactobacillus kefir and Saccharomyces lipolytica isolated from kefir grains: evidence for lectin-like activity of bacterial surface proteins. J Dairy Res 76:111–116CrossRefPubMedGoogle Scholar
  11. Guo Z, Wang J, Yan L, Chen W, Liu X-M, Zhang H-P (2009) In vitro comparison of probiotic properties of Lactobacillus casei Zhang, a potential new probiotic, with selected probiotic strains. LWT-Food Sci Technol 42:1640–1646CrossRefGoogle Scholar
  12. Hollmann A, Delfederico L, Glikmann G, De Antoni G, Semorile L, Disalvo E (2007) Characterization of liposomes coated with S-layer proteins from lactobacilli. Biochim Biophys Acta 1768:393–400CrossRefPubMedGoogle Scholar
  13. Jakava-Viljanen M, Palva A (2007) Isolation of surface (S) layer protein carrying Lactobacillus species from porcine intestine and faeces and characterization of their adhesion properties to different host tissues. Vet Microbiol 124:264–273CrossRefPubMedGoogle Scholar
  14. Jankovic I, Ventura M, Meylan V, Rouvet M, Elli M, Zink R (2003) Contribution of aggregation-promoting factor to maintenance of cell shape in Lactobacillus gasseri 4B2. J Bacteriol 185:3288–3296CrossRefPubMedPubMedCentralGoogle Scholar
  15. Johnson B, Selle K, O’Flaherty S, Goh YJ, Klaenhammer T (2013) Identification of extracellular surface-layer associated proteins in Lactobacillus acidophilus NCFM. Microbiology 159:2269–2282CrossRefPubMedPubMedCentralGoogle Scholar
  16. Khaleghi M, Kermanshahi RK, Yaghoobi M, Zarkesh-Esfahani S, Baghizadeh A (2010) Assessment of bile salt effects on s-layer production, slp gene expression and some physicochemical properties of Lactobacillus acidophilus ATCC 4356. J Microbiol Biotechnol 20:749–756PubMedGoogle Scholar
  17. Kim WS, Perl L, Park JH, Tandianus JE, Dunn NW (2001) Assessment of stress response of the probiotic Lactobacillus acidophilus. Curr Microbiol 43:346–350CrossRefPubMedGoogle Scholar
  18. Kos B, Šušković J, Vuković S, Šimpraga M, Frece J, Matošić S (2003) Adhesion and aggregation ability of probiotic strain Lactobacillus acidophilus M92. J Appl Microbiol 94:981–987CrossRefPubMedGoogle Scholar
  19. Lee Y-K, Puong K-Y, Ouwehand AC, Salminen S (2003) Displacement of bacterial pathogens from mucus and Caco-2 cell surface by lactobacilli. J Med Microbiol 52:925–930CrossRefPubMedGoogle Scholar
  20. Leeuw E, Li X, Lu W (2006) Binding characteristics of the Lactobacillus brevis ATCC 8287 surface layer to extracellular matrix proteins. FEMS Microbiol Lett 260:210–215CrossRefPubMedGoogle Scholar
  21. Lorca G, Torino MI, Font de Valdez G, Ljungh Å (2002) Lactobacilli express cell surface proteins which mediate binding of immobilized collagen and fibronectin. FEMS Microbiol Lett 206:31–37CrossRefPubMedGoogle Scholar
  22. Lu R-R, Qian P, Sun Z, Zhou X-H, Chen T-P, He J-F, Zhang H, Wu J (2010) Hempseed protein derived antioxidative peptides: purification, identification and protection from hydrogen peroxide-induced apoptosis in PC12 cells. Food Chem 123:1210–1218CrossRefGoogle Scholar
  23. Martínez MG, Acosta MP, Candurra NA, Ruzal SM (2012) S-layer proteins of Lactobacillus acidophilus inhibits JUNV infection. Biochem Biophys Res Commun 422:590–595CrossRefPubMedGoogle Scholar
  24. Meng J, Zhu X, Gao S-M, Zhang Q-X, Sun Z, Lu R-R (2014) Characterization of surface layer proteins and its role in probiotic properties of three Lactobacillus strains. Int J Biol Macromol 65:110–114CrossRefPubMedGoogle Scholar
  25. Meng J, Gao S-M, Zhang Q-X, Lu R-R (2015) Murein hydrolase activity of surface layer proteins from Lactobacillus acidophilus against Escherichia coli. Int J Biol Macromol 79:527–532CrossRefPubMedGoogle Scholar
  26. Mobili P, Londero A, Maria TMR, Eusébio MES, De Antoni GL, Fausto R, Gómez-Zavaglia A (2009) Characterization of S-layer proteins of Lactobacillus by FTIR spectroscopy and differential scanning calorimetry. Vib Spectrosc 50:68–77CrossRefGoogle Scholar
  27. Navarre WW, Schneewind O (1999) Surface proteins of gram-positive bacteria and mechanisms of their targeting to the cell wall envelope. Microbiol Mol Biol Rev 63:174–229PubMedPubMedCentralGoogle Scholar
  28. Sára M, Sleytr UB (2000) S-layer proteins. J Bacteriol 182:859–868CrossRefPubMedPubMedCentralGoogle Scholar
  29. Sleytr UB (1997) Basic and applied S-layer research: an overview. FEMS Microbiol Rev 20:5–12CrossRefGoogle Scholar
  30. Smit E, Pouwels PH (2002) One repeat of the cell wall binding domain is sufficient for anchoring the Lactobacillus acidophilus surface layer protein. J Bacteriol 184:4617–4619CrossRefPubMedPubMedCentralGoogle Scholar
  31. Sun Z, Huang L, Kong J, Hu S, Zhang X, Kong W (2012) In vitro evaluation of Lactobacillus crispatus K313 and K243: high-adhesion activity and anti-inflammatory effect on Salmonella braenderup infected intestinal epithelial cell. Vet Microbiol 159:212–220CrossRefPubMedGoogle Scholar
  32. Ventura M, Jankovic I, Walker DC, Pridmore RD, Zink R (2002) Identification and characterization of novel surface proteins in Lactobacillus johnsonii and Lactobacillus gasseri. Appl Environ Microbiol 68:6172–6181CrossRefPubMedPubMedCentralGoogle Scholar
  33. Zhang Y-C, Zhang L-W, Tuo Y-F, Guo C-F, Yi H-X, Li J-Y, Han X, Du M (2010) Inhibition of Shigella sonnei adherence to HT-29 cells by lactobacilli from Chinese fermented food and preliminary characterization of S-layer protein involvement. Res Microbiol 161:667–672CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature and the University of Milan 2018

Authors and Affiliations

  1. 1.School of Food Science and Technology, Province Key Laboratory of Transformation and Utilization of Cereal ResourceHenan University of TechnologyZhengzhouChina
  2. 2.School of Food Science and TechnologyJiangnan UniversityWuxiChina

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